Method for Estimating RNA Integrity

An automated method for estimating the integrity of RNA samples based on microcapillary electrophoretic RNA separation has been developed.

The RNA integrity number (RIN) is a software tool designed to help scientists estimate the integrity of RNA samples. The RIN extension automatically assigns an integrity number to a eukaryote total RNA sample. Using this tool, sample integrity is determined by the entire electrophoretic trace of the RNA sample. This includes the presence or absence of degradation products.

The Agilent (Palo Alto, CA, USA) 2100 bioanalyzer is a bioanalytic device based on a combination of microfluidics, microcapillary electrophoresis, and fluorescence detection, which provides a platform to record the size and distribution of RNA molecules in a digital format. The approach is automated, user independent, and reproducible, and enables evaluation of the integrity of RNA samples using a software logarithm.

The RIN can be used for a number of purposes: to directly compare RNA samples, e.g., before and after shipment, to compare the integrity of the same tissue across different laboratories, to ensure the repeatability of experiments. For example, if RIN shows a given value and is suitable for microarray experiments, then RIN of the same value can be always used for similar experiments if the same organism, tissue, and/or extraction method is used.

In the January 31, 2006, issue of BiomedCentral (BMC) Molecular Biology, Odilo Mueller and colleagues at Agilent describe how a large collection of electrophoretic RNA measurements recorded with the Agilent 2100 bioanalyzer was used to extract an algorithm that describes RNA integrity. Their results show the importance of taking characteristics of several regions of the recorded electropherogram into account to obtain a robust and reliable prediction of RNA integrity.



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